TP53 Rabbit Polyclonal Antibody

25 ug of the indicated Human whole cell extracts was loaded onto a 12% SDS-PAGE gel. 1 ug/ml of the antibody was used in this experiment. TP53 is expressed as several different isoforms (> 10) ranging in size from 24 kDa to ~50 kDa in a cell type specific manner. This immunogen for this antibody is present in several isoforms. 1-3 ug/ml is recommended for this antibody.

DU145 cells were lysed in IP lysis buffer (20mM HEPES, 1% Triton X-100, 150mM NaCl, 1mMEDTA, 1mM EGTA, 100mM NaF, 10mM Na4P2O7, 1mM Na3VO4, 0.2mM PMSF). Amount of protein per well: 30 ug, Primary antibody conditions: 1:2000 in 5% milk/TBST buffer, overnight at 4°C.Secondary antibody conditions: 1:5000 in 5% milk/TBST buffer, 1 hour at room temperature. TP53 is strongly supported by BioGPS gene expression data to be expressed in Human DU145 cells.

Sample type: HeLa cells Antibody titration: 1 to 1000, Gel: 4-15% gradient, Secondary: Goat anti-rabbit HRP, Secondary dilution: 1 to 5000. TP53 is supported by BioGPS gene expression data to be expressed in HeLa.

U20S (p53+) cells were treated with 0.5 uM Doxorubicin for 14 hrs to induce DNA damage and hence activate p53. In parallel, PLKO cells (U2OS cells with stable shRNA-mediated knockdown of p53) were treated similarly and were used as negative control. Thedata for p21 promoter were normalised to actin (control for non-specific binding of DNA to the antibodies).

WB Suggested Anti-TP53 Antibody Titration: 0.2-1 ug/ml, ELISA Titer: 1:1562500, Positive Control: 293T. TP53 is strongly supported by BioGPS gene expression data to be expressed in HEK293T.